Method of treating malignancies and viral infections and improving immune function with a dietary supplement

ABSTRACT

A dietary supplement comprising as its active ingredients soybean, mushroom and mung bean is used in a method to in a method to ameliorate at least one effective malignancy or viral infection or to enhance immune functions. The active components of the dietary supplement include phytoestrogens, β-glucans, saponins, inositol hexaphosphate, and lectins.

TECHNICAL FIELD

[0001] This invention relates to treatments effective in treatingmalignancies and restoring CD4⁺ cell levels, and also to dietarysupplements that improve immune functions. In particular, it concernscompositions that are derived from plants, foods and extracts thereof,that are used in a particular combination as dietary supplements, whichare effective against malignancies and depressed immune functions andwhich improve overall immune function.

BACKGROUND OF THE INVENTION

[0002] Nutritional support for cancer patients has been studiedextensively. It provided a substantial improvement in nutritional statusof cancer patients whose malnutrition was due to reduced oral intake,but it fell short of its goals when the malnutrition was due tomalignancy. No improvement in the tumor's response to therapy has beendemonstrated by this approach, which primarily manipulated protein andcaloric intake in a generic strategy.

[0003] In the past three decades, studies have repeatedly shown thatsome foods contain components with antitumor or immune-enhancingactivities. These components are nontoxic and have different modes ofaction, and their individual antitumor activities are modest. A partiallist of these components is presented in Table 1. The use of fruits andvegetables, which were not specifically selected for their antitumorattributes and may contain only a few of these components, for cancerprevention has been recommended by the American Cancer Society and theNational Cancer Institute; their increased consumption is considered amajor cause for the reduction in cancer-related deaths in recent years.TABLE 1 Non-toxic Antitumor Components in Plants¹ Antitumor ComponentsFunctions Protease inhibitors in soybean Suppress chemically inducedcarcinogenesis in colon, anal gland, oral cavity, lung, breast, and skinAutoclave-resistant factor in Protection against metastasis ofradiation-induced soybean thymic lymphosarcoma and weight loss Plantsterols Protective effect against tumorigenesis in colonic mucosaSaponin (found in soybeans) Potentiation of rabies vaccine in miceAntitumor activity Inhibition of induction of Epstein-Barr virusGenistein (found in soybeans) Inhibits angiogenesis and growth of humanprostate cancer Biochanin A (found in soybeans & Inhibits human prostatecancer mung beans) Other isoflavones Inhibit growth of human cancer celllines derived from gastrointestinal tract Inositol hexaphosphate (foundin Enhances NK cell activity and suppresses tumor soybeans or mung bean)growth Suppresses tumor formation and growth Inhibits growth of humanleukemia cells and induces differentiation of K-562 cells β-Glucans(e.g., lentinan) (found in Host-dependent inhibition of sarcoma 180 cellmushrooms) growth Immunoaccelerators, immunopotentiators, andimmunomodulators Enhance production of IL-1, TNF, LAK activity,cytotoxic T lymphocytes, cytotoxic peritoneal exudate cells Lectins(found in mung beans) Selective stimulation of T lymphocyteproliferation

[0004] Non-small cell lung cancer (NSCLC) causes the death of >400,000patients annually in the United States and Europe. The benefit ofconventional therapies for unresectable stage IIIB and IV NSCLC patientsis marginal; the generally accepted median survival time (MST) of stageIV patients has remained 4-6 months for the control group and 6-10months for the treatment group, and the one-year survival time isusually only 20%. Even stage I NSCLC patients have a 52% death rate dueto lung cancer within two years when treated with radiotherapy alone.Clearly, more effective therapies are needed.

[0005] With regard to AIDS patients, improvement in CD4⁺ cell count hasbecome a major area of research in AIDS therapies. Current therapies cannot increase CD4⁺ counts to normal range in the 12-month treatmentperiod. Many patients are still dying from depressed immune functionseven when their serum HIV load can be reduced to an undetectable levelby antiretroviral therapy.

[0006] Continuous treatment with toxic antiretroviral drugs for a longperiod of time has generated instances of intolerable toxicity; manypatients cannot continue these highly toxic treatments. Therefore, itwould be useful to the industry to provide non-toxic treatments that areeffective against malignancies and viral infections.

DISCLOSURE OF THE INVENTION

[0007] The invention is directed to a dietary supplement and a protocolof administration that is used to treat malignancies and viralinfections and to otherwise improve overall immune function. The dietarysupplement is a selected vegetable (SV) composition that is afreeze-dried commercial vegetable soup prepared by Sun Farm Corporation(Milford, Conn.). It is a brown powder with a slightly sweet taste. Theingredients of SV include soybean, mushroom, mung bean, red date,scallion, garlic, lentil bean, leek, hawthorn fruit, onion, ginseng,angelica root, licorice, dandelion root, senegal root, ginger, olive,sesame seed, and parsley. Rather than focusing on a single immunemodulator, SV is a combination of various kinds of herbs and vegetablesselected for their specific activities. Thus, immune function may beaugmented in a concerted effort.

BRIEF DESCRIPTION OF THE DRAWING

[0008]FIGS. 1a and 1 b are graphs illustrating the growth of lung tumorsin BALB/c mice.

[0009]FIG. 2 is a CT scan of three metastatic brain lesions illustratingtheir disappearances after the SV treatment.

[0010]FIG. 3 shows the median survival time of Stage III B and IVnon-small cell lung cancer patients in the pilot study.

[0011]FIG. 4a shows the right cerebellar metastasis of patient #9 in thepilot study.

[0012]FIG. 4b is a radiograph of a left scapula of patient #8 in thepilot study.

[0013]FIG. 5 illustrates the survival times of stage III and IVnon-small cell lung cancer patients with and without the selectedvegetable treatment.

MODES OF CARRYING OUT THE INVENTION

[0014] The combination of SV as further described contains soybean,mushroom, and mung bean as the active components. The remainder of thecomponents, red date, scallion, garlic, lentil bean, leek, hawthornfruit, onion, ginseng, angelica root, licorice, dandelion root, senegalroot, ginger, olive, sesame seed, and parsley are added for flavor. Themain active components of soybean, mushroom and mung bean, aregenestein, biochanin A, coumestrol, inositol hexaphosphate, β-glucans,and lectins.

[0015] Vegetables and herbs were selected for inclusion in SV on thebasis of two criteria: absence of toxicity and presence of one or moreknown antitumor or immune-enhancing components as found in Table 1.

[0016] One embodiment of the invention is a method to ameliorate atleast one effect of malignancy or viral infection comprisingadministering to a subject in need of such treatment an effective amountof the dietary supplement comprising soybean, shiitake mushroom, andmung bean, wherein the composition is administered periodically for atime to achieve amelioration.

[0017] Another embodiment of the invention is a method of enhancingimmune functions comprising administering to a subject an effectiveamount of a dietary supplement comprising soybean, shiitake mushroom andmung bean periodically for a time to achieve enhanced immune functions.

[0018] Anti-viral effects such as increasing CD4⁺ cell levels, have beenfound in patients from HIV, Epstein-barr virus, herpes, and influenzaviruses. In addition, malignancies such as small cell lung cancer,non-small cell lung cancer, prostate cancer, breast cancer, stomachcancer, small intestine cancer, colonrectal cancer, renal cellcarcinoma, soft tissue sarcoma, ovarian cancer, thyroid cancer,acrcinoid tumors, sarcoma of the bone, melanoma, glioblastoma,oligodendroglioma, astrocytoma, medulloblastoma, acute lymphomas,chronic leukemias, cancer of unknown primary site, Hodgkin's disease,and non-Hodgkin's disease have been successfully treated with anadministration of the dietary supplement in accordance with theinvention. Moreover, as this dietary supplement is non-toxic and hasimmune enhancing properties, it can be used to preventimmuno-suppressing diseases.

[0019] In general, the amounts of material administered are in the rangeof those corresponding to 25-200 gms. of the soybean, shiitake mushroom,and mung bean components and similar amounts of the optional additives.

[0020] The dietary supplement can be in the form of a freeze-driedproduct or extracts of the essential components along with theflavorings. The freeze-dried material or extracts can be added to wateror other soup and administered orally on a regular basis. SV can beboiled with water or soup and frozen for convenience and then heated foruse. About 10-50 mg per 50 kg body weight of the freeze dried or netweight SV is preferable and about 30 mg per 50 kg body weight is morepreferable. Thus, about 100-500 mg of about a 10% SV mixture with aliquid per 50 kg body weight is preferable and about 300 mg is morepreferable. Preferably, the dietary supplement should be administeredonce or twice daily. Administration of 3 to 4 times a week is alsoacceptable.

[0021] In another embodiment, the present invention is directed to thedaily ingestion of nontoxic vegetables and herbs containing knownantitumor components that can improve clinical outcomes of patients whoalready have advanced tumors. These observations suggest the benefits ofSV and led to the present invention.

[0022] Various species of mushroom can be used in the regimen of theinvention. Acceptable species include: Polyponis frondosus; Polyporusgiganteus; Polyporus umbellatus; Trametes cinnabarina. However, thefollowing are more preferable: Armillariella mellea; Armillariellatabescens; Clitocybe nebularis; Lentinus lepideus Fr.; Panus rudis Fr.;Pleurotus spodolencus Fr.; Pholiota adiposa; Boletus erythropus; andGyroponis castaneus. More preferred are: Flammulina velutipes; Lepistaluscina (Fr) Sing.; Lepista nuda (Bull. ex. Fr.); Oudemansiellaradicata; Tricholomopsis platyphylla; Panellus stypticus; Pleurotusostreatus; Schizophyllum commune Fr.; Tricholoma gambosum, Agaricusarvensis; Coprinus atramentarius; Coprinus comatus; Agrocybe palidosa;Agrocybe praecox; Pholiota lubrica; Pholiota nameko; Rozites caperata;Cortinarius bovinus Fr.; Cortinarius salor Fr.; Cortinarius violaceus;Gymnopilis liquiritae; Rhodophyllus clypeatus; Rhodophyllus sinuatus;Boletus edulis Bull.; Boletus pulvertlentus; Boletus regius Krombh.;Boletus rubellus Krombh.; Suillus aeruginascens; Suillus bovinus;Suillus liteus; Lactarius hygrophoroides; Lactarius volemus Fr.; Russulaadusta Fr.; Russula crustosa; Russula delica Fr.; and Russula lepida Fr.Especially preferred are: Lentinus edodes; Panus conchatus; Tricholomamatsutake; Tricholoma monolicum Imai; Agaricus bisporus; Agaricuscampestris; Cortinarius cinnamomeus Fr.; Cortinarius collinitus Fr.;Cortinarius elatior Fr.; Cortinarius latus Fr.; and Lactarius hatsudake.

[0023] A table of the properties of the various species follows: PercentInhibition of Species Name Edible Tumor Growth Polyporus frondosus yes  98% Polyporus giganteus yes   90% Polyporus umbellatus yes   70%Trametes cinnabarina yes   90% Armillariella mellea yes   80% cureophthalmocopia Armillariella tabescens yes   70% Clitocybe nebularis yes  80% Flammulina velutipes good 80-100% Lentinus edodes excellent 80-97% Lentinus lepideus Fr. ywa  50-70% Lepista luscina (Fr.) Sing.delicious yes Lepista nuda (Bull.) ex. Fr.) yes 90-100% Oudemansiellaradicata Tricholomopsis platyphylla yes  80-90% Panellus stypticus yes 70-80% Panus conchatus yes   100% Panus rudis Fr. yes   70% Pleurotusostreatus yes   80% Pleurotus spodolencus Fr. yes   72% Schizophyllumcommune Fr. yes   100% Tricholoma gambosum yes  70-90% Tricholomamatsutake excellent 70-100% Tricholoma mongolicum Imai excellent 70-100%Agaricus arvensis yes   100% Agaricus bisporus excellent   100% Agaricuscampestris excellent   100% Coprinus atramentarius yes   100% Coprinuscomatus yes   100% Agrocybe paludosa yes   100% Agrocybe praecox yes  100% Pholiota adiposa yes  70-90% Pholiota lubrica yes   100% Pholiotanameko yes   100% Rozites caperata yes   70% Cortinarius bovinus Fr. yes  90% Cortinarius collinitus Fr. yes   100% Cortinarius elatior Fr. yes  80% Cortinarius latus Fr. yes   100% Cortinarius salor Fr. yes   90%Cortinarius violaceus yes   100% Gymnopilus liquiritae yes   90%Rhodophyllus clypeatus yes   100% Rhodophyllus sinuatus yes   100%Boletus edulis Bull, yes 90-100% Boletus erythropus ?   100% Boletuspulverulentus yes   90% Boletus regius Krombh. yes   90% Boletusrubellus Krombh. yes   90% Gyroporus castaneus yes   80% Suillusaeruginascens yes   100% Suillus bovinus yes   100% Suillus luteus yes  90% Lactarius hatsudake yes   100% Lactarius hygrophoroides yes   70%Lactarius volernus Fr. yes   90% Russula adusta Fr. yes   80% Russulacrustosa yes   70% Russula delica Fr. yes   100% Russula lepida Fr. yes  100%

[0024] The active components in the SV comprise phytoestrogens,β-glucans, saponin, inositol hexaphosphate, and lectins. Phytoestrogenssuch as genistein, coumestrol, daidzein, and biochanin A are eachpresent in the SV based on daily oral consumption of about 1 mg-5 g/100kg of body weight. β-glucans such as lentinan, pachymaran, zymosan,schizophyllan, KS-2, and pachyman are each present in SV based on adaily oral consumption in an amount of about 1 mg-5 g/1100 kg of bodyweight. Saponins and inositol hexaphosphate are each present in the SVbased on daily oral consumption of about 1 mg-5 g/100 kg of body weight.Lectins, such as phytohemagglutinin and concanavalin A are each presentin the SV based on daily oral consumption of from about 1 mg-5 g/10 kgof body weight.

[0025] The following examples serve to illustrate, but not to limit, theinvention.

[0026] The first example that follows illustrates the influence ofselected vegetables (SV) that contain known antitumor components on thesurvival of stage III-IV non-small cell lung cancer (NSCLC) patients.All patients were treated with conventional therapies. SV was added tothe daily diet of 5 stage I patients in the toxicity study group (TG)and 6 stage III and IV patients in the treatment group (SVG), but not tothe diet of 13 stage III and IV patients in the control group (CG). Age,Karnofsky performance status (KPS), and body mass index of SVG and CGpatients were comparable at entry. KPS declined in the CG patients (79±8to 55±11) but improved in the SVG patients (75±8 to 80±13) one to threemonths after entry. Weight change in the CG, SVG, and TG patients was−12±5%, −2±2%, and +4±4%, respectively. The median survival time andmean survival of the CG patients were 4 and 4.8 months, but in the SVGpatients they were 15.5 and 15 months (p<0.01). No clinical signs oftoxicity were found in the TG patients in the 24-month study period.Adding SV to the daily diet of NSCLC patients was found to be nontoxicand associated with improved weight maintenance, KPS, and survival ofstage III and IV NSCLC patients.

EXAMPLE 1 Pilot Study of a Selected Vegetable Diet in Tumor Bearing Miceand Stage IIIB and IV Non-Small Cell Lung Cancer Patients. Method

[0027] SV Preparation. SV was prepared by Sun Farm Corporation, Milford,Conn., employing “The Good Manufacturing Practices” designed toguarantee minimal contamination with heavy metals and bacteria (U.S.Federal Code of Regulations, Title 21, Part 100-69. 1996; U.S.Government Printing Office, Washington, D.C.). The ingredients of SVinclude: soybean, mushroom, mung bean, red date, scallion, garlic,lentil bean, leek, Hawthorn fruit, onion, ginseng, angelica root,licorice, dandelion root, senegal root, ginger, olive, sesame seed, andparsley. The mixture was boiled and then stored frozen at −20° C. andthawed at room temperature before use.

[0028] Nutritional Data and Toxicity. Random samples of SV were sent toNortheast Laboratory, Berlin, Conn., for analysis of nutritional value,heavy metals, and bacteria.

[0029] Phytoestrogens: The assay was modified from Franke et al. (FrankeA A, Custer L J, Cema C M, Narala K K. Quantitation of phytoestrogens inlegumes by HPLC. J. Agric Food Chem 1994; 42: 1905-13). One gram offreeze-dried SV (DSV) was dispersed in a 50 ml mixture ofHCl/ethanol/butylated hydroxytoluene (10/40/0.25, v/v/v), refluxed for 2h, filtered, and 5 μl of the filtrate was analyzed with HPLC (Nova-Pak150 mm×3.9 mm reverse phase C18 column, Waters, Milford, Mass.). Thecolumn was eluted with a gradient from 5% to 50% acetonitrile in aceticacid-water (1/9, v/v) in 15 min at 1 ml/min followed with 7.5 min washwith 50% acetonitrile in the same buffer. Molar extinction coefficients(daidzein, 20,893 at 250 nm; genistein, 37,154 at 263 nm, coumestrol,22,300 at 339 nm) were used for quantitative determination.

[0030] Inositol: DSV (5 g) was incubated with 10 ml of 0.33N HCl for 2h, filtered, and diluted with deionized water (1:1, v/v). Inositolhexaphosphate was determined according to Ellis et al. (Ellis R. MorrisE R, and Philpot C. Quantitative determination of phytate in thepresence of high inorganic phosphate. Anal Biochem 1977; 77:536-9.)

[0031] Mouse Lung Tumor Model: Four week-old BALB/c male mice weredivided into groups of 5 (Experiment I) or 8 (Experiment II) mice. Micein each group were fed one food preparation a week prior to tumorinoculation and during the entire study period.

[0032] BALB/c Line 1 Lung tumor cells (10⁴ cells; viability over 90%)were injected subcutaneously in the right thigh. Tumor size was measuredevery 2-4 days. Experiment I: Food #1 was made from Lab Chow powder,mixed with water, made into pellets, and air-dried. Food #2, #3, and #4were made as above and mixed with a hot water extract of mung bean (#2:10% by wt; Lab Chow, 90%), shiitake mushroom (#3: Lentinus edodes, 10%by wt), or both at the same concentrations (#4). Experiment II: Lab Chowpowder (#5, Control) was mixed with freeze-dried SV (DSV) powder (#6: 5%wt/wt). Food consumption per group was measured weekly.

[0033] Clinical Study Design. Sixteen patients with knowledge of thelead cases requested SV as a nutritional supplement and volunteeredtheir participation. The study period was 60 months from February, 1992to January, 1997. Patients were treated with conventional therapiesselected by their physicians and added SV to their daily diet. Theirclinical status was monitored by their physicians and verified by theauthors. Only 14 stage IIIB and IV patients who ingested SV daily for 2months or longer were included in the study group. Kamofsky performancestatus (KPS) was recorded before the use of SV and 5 months later. Thefour patients who ingested SV less than 2 months were excluded fromanalyses. Patients #1 and #2 provided retrospective data; they wereexcluded from the statistical analyses.

[0034] Diagnosis, Staging, and Survival Time. All the patients werediagnosed in their local hospitals. The pathological slides werereviewed by the pathologist (TMF) and radiographs by the radiologists(YPH and HCY). Staging was determined according to the revisedInternational System for Staging Lung Cancer. The survival time wascalculated from the date of diagnosis of stage IIIB or IV NSCLC to thedate of death or to the cut-off date.

[0035] Ingestion of SV. SV is a grayish green paste with a slightlysweet taste. It was ingested daily (approximately 283 g) by patients aspart of breakfast or lunch either directly, or mixed with other foods.

[0036] Materials: All chemicals were reagent grade. Genistein, daidzein,inositol, inositol hexaphosphate, butylated hydroxytoluene, andO-toluidine were obtained from Sigma Aldrich Chemical Co. (St. Louis,Mo.), coumestrol, acetonitrile, acetic acid, trichloroacetic acid, andthiourea from Fisher Scientific (Pittsburgh, Pa.). Purina Lab Chowpowder from Purina Mills (Richmond, Ind.), BALB/c mice from CharlesRiver Laboratories (Wilmington, Mass.), and SV from Sun Farm Corporation(Milford, Conn.).

[0037] Informed Consent: All patients were informed of the ingredientsin SV and that these ingredients have been used widely as foods or foodflavors. They used SV daily as a food supplement, and gave writtenpermission to the authors to review their medical records.

Results

[0038] Nutritional Value and Hazardous Contaminants of SV. The dailydose of SV contains a net wet weight of 283 grams; physiological fuelcalories, 87+3; protein, 10+2 g; carbohydrate, 11+2 g; total fat, <1 mg;cholesterol, <1 mg; dietary fiber, 12+1 g; vitamin C<0.1 mg; vitamin A,<15 ITJ; thiamine, <4 mg; riboflavin, 45+4 mg; sodium, 45+4 mg; calcium,9+1 mg; potassium, <1 mg; iron, <5 mg. Heavy metal content and cultureplate counts of samples of SV were: arsenic <1 (PPM); cadmium, <1.3;barium, <1; chromium, <2.7; lead, <1.7; mercury, <0.1; silver, <1.3;yeast and mold count (per gram), <10; E. coli, <1; and standard platecount, <10. These levels are well below allowable levels.

[0039] Antitumor Components in SV: The following components weremeasured in SV (mg/283 g SV/daily dosage): inositol hexaphosphate, 63+6;genistein, 2.6+0.1; daidzein, 4.4+0.1; coumestrol, 15.5+0.4.

[0040] Mouse Lung Tumor Model: In Experiment I, tumor size was measuredon the 10th, 12th, 14th, 17th, and 22nd day after the inoculation oftumor cells (FIG. 1A). Curves #1, #2, #3, and #4 in FIG. 1A depict themean tumor size in mice consuming Food #1, #2, #3, and #4 respectivelymeasured after inoculation. Each point is mean±SE. The percentinhibition of tumor growth rate in groups #2, #3, and #4 compared withthe control (#1) were 16%, 49%, and 82% on the 14th day and 53%, 60%,and 82% on the 22nd day respectively. Both mung bean (#2) and mushroomextract (#3) showed clear inhibition of tumor growth and these effectswere additive when both extracts (#4) were used in combination. InExperiment II, tumor size was measured on the 14th 16th, 19th, 21st, and23rd day after tumor inoculation (FIG. 1B). Curves #5 and #6 in FIG. 1Bare the mean tumor size in mice using Food #5 and #6. Each point ismean±SE. Lab Chow containing 5% DSV produced 53-74% inhibition in tumorgrowth rate in the first 23rd days. One of 8 mice showed partial tumorregression after 26th day and complete regression after 32 days. Theweekly food consumption per mouse of preparations #1 to #6 was 16.9+1.4g, 18.9+0.8 g, and 16.0+0.6 g in the first, second, and third week aftertumor inoculation. No appreciable difference between groups was found.

[0041] Patient Characteristics: The patients' gender, age, stage, celltype, metastatic sites, conventional treatments to the primary tumor andmetastatic lesions respectively, tumor response, complications, and KPSare summarized in Table 2. Occurrence of metastatic disease afteringesting SV, the length of time taking SV, the duration of tumor-freestatus, and survival are also summarized in Table 2. TABLE 2 PatientsCharacteristics and Survival Time with Adjuvant SV to Therapy TreatmentsOther New T Time (Months)*³ Tumor Stage Cell Metastatic PrimaryMetastatic Tumor Compli- after KPS*² using Case/Sex/Age T.N.M. Type SiteTumor Tumors Response*¹ cations SV (a) (b) SV Free Survival A. LeadCases:  1 F 69 IV: 2.2.1. lrg adrenal S C, R, SV, no residual T no no 40100 27 >133 >137 smancs, S alive  2 F 75 IV: 2.1.1 adn brain S SV, R,complete mal- no 40 80 12 10 14 smancs nutrition^(P2) B. Study Group a.Complete Response of Metastatic Tumors and No Recurrent Tumor AfterUsing SV Alone  3 F 57 IV: 1.3.1. adn both lungs S SV complete no no 50100 >35 >30 >39 alive  4 F 73 IV: 1.3.1. adn both lungs S SV complete nono 90 100 >27 >12 >28 alive b. Complete Response, Partial Response, orStabilization of Metastatic Tumor after Using SV and Other Treatments  5F 80 IV: 4.2.1. adn vert, pleu R R, SV stable/prog no no 50 90 >46 0 >48ef alive  6 M 67 IIIB: 4.2.0. adn pleural eff no PD, SV, partial nerveyes 50 90 43 0 45 R, C injury^(P6)  7 F 34 IIIB: 2.3.0. nsc contral l.n.S C, SV, R, no residual T no no 50 100 >32 >30 >33 alive  8 F 78 IIIB:4.0.0. adn pleural eff S R, C, SV stable pneumonia no 50 100 >21 >20 >28alive  9 M 69 IV: 2.0.1. adn brain, S R, SV complete no yes 50 100 16 na22 bone 10 F 51 IV: 1.2.1. adn brain S, C, R SV, R, S no residual Tdepres- yes 50 90 14 na 20 sion^(P10) 11 F 59 IV: 1.1.1. adn brain noSV, R stable/prog v. no 50 90 10 0 11 throm- boses^(P11) 12 M 58 IV:2.3.1. adn adrenal S, C C, SV, C stable/prog heart^(P12) no 60 70 9 0 1213 F 76 IIIB: 4.3.0. adn pleural eff S C, R, SV complete no no 50 100 52 >8 alive 14 F 47 IIIB: 4.3.0. adn pleural eff C C, SV na na 50 50 5 06  Average Karnofsky Performance Status 54 ± 12 90 ± 15  (Pts #3-#14) C.Ineligible Patients*⁴ 15 M 40 IIIA: 1.2.0. adn C, R SV nausea, vomiting1.5 0 10 16 M 56 IV: 4.0.1. sqm vert R R, SV paraplegia 1.4 0 5 17 M 64IV: 4.0.1. adn brain, S, C SV 0.2 0 6 bone 18 M 57 IV: 4.0.1. adn spine,rib C, R SV 0.5 0 3

[0042] Lead Cases:

[0043] Case #1: A stage IV patient whose tumor progressed while onchemotherapy and radiotherapy has been tumor-free for 133 months afteringestion of SV, SMANCS therapy, and resection of a metastatic lesion.

[0044] Patient #1 had a poorly differentiated large cell carcinoma inher RLL (2.2×2 cm) with a positive subcarinal lymph node (Table 2). Alobectomy in 1/85 was followed with methotrexate, adriamycin, CCNU, andcytoxan. Her disease progressed with metastasis to left adrenal gland(4×3 cm, 8/85), a right pleural effusion developed, and her KPS droppedto 40. Her adrenal tumor did not respond to radiotherapy and continuedgrowing (5×4 cm, 11/85). She began ingesting SV daily from 9/85. She wasalso treated with SMANCS, which is the antitumor protein,neocarcinostatin, conjugated with a polystyrene-maleic acid copolymer(35), and had adrenalectomy in 12/85. The adrenal tumor was wellencapsulated containing mostly necrotic tissue. She continued SMANCS for4 months and ingested SV daily until 12/87. She remains tumor-free.

[0045] Case #2: Complete regression of three metastatic brain tumors

[0046] Patient #2 had a well differentiated adenocarcinoma in RUL (Table2). She had a lobectomy in 3/86. In 10/86, CT scan showed 3 metastaticbrain lesions (FIG. 2). KPS was 40. She started SV, SMANCS, and a onemonth course of radiotherapy. All 3 tumors disappeared completely in 4months. She continued SV and SMANCS therapy until 6/87. During this timeshe developed anorexia and severe malnutrition, followed by aspirationpneumonia and sepsis, and died in 12/87 without clinical signs of tumorrecurrence.

[0047] Median Survival Time: Five stage IIIB and 7 stage 1V patientswere eligible for analysis (Patients #3-14). Half of these patients werestill alive at the closing date; thus, the MST was not reached accordingto Kaplan and Meier (Kaplan EL, Meier P. Nonparametric estimation fromincomplete observations. J Am Stat Assoc 1958; 53: 457-82). Aconservative estimate of 33.5 months, namely, the midpoint of thesurvival curve above the median, i.e., between 22 and 45 months, waschosen as the MST (FIG. 3). As shown in FIG. 3, the survival of 12patients (#3 to #14, Table 2) was plotted and analyzed according toKaplan and Meier. The MST should be 45 months and the 95% confidenceinterval had a lower boundary of 12 months and no upper boundaryreached. For conservative evaluation, the midpoint of the survival curveabove the median was chosen as MST, namely, between 22 and 45 months or33.5 months. The 95% confidence interval had a lower-boundary of 12months and no upper-boundary. One-year survival was 75%.

[0048] New Tumors: Ten of the 14 patients (#1-5, 7, 8, 11-13, Table 2)had no new sites of metastasis after conventional treatments and SVtherapy. Patient #14 suffered severe side-effects of concurrentchemotherapy and was not evaluated for new tumors.

[0049] SV Therapy Alone: Both patient #3 and #4 had complete regressionof their multiple metastatic lesions in both lungs after using SV alonefor 5 and 15 months and remained tumor-free, 39 and 28 months afterdiagnosis. After unsuccessful radiotherapy and while using SV alone, thepulmonary and vertebral body lesions of patient #5 were stable for 40months and subsequently progressed slowly. She was still alive at theconclusion of this study.

[0050] SV and Radiotherapy: Of the four patients who had brainmetastases, three (#2, #9, 10) achieved complete regression after usingSV and radiotherapy. Both lesions in brain and bone of Patient #9regressed completely after radiotherapy. FIG. 4 is a right cerebellarmetastasis of patient 9. FIG. 4A shows in A is a post contrast CT scanin superior orbitomeatal plane taken on Dec. 17, 1992. An enhancingmetastatic lesion (T) of 1.5×1.2× approximately 1.2 cm in size, locatedin the quadrangular lobule of the cerebellum on the right side withconsiderable edema (low density area) of the surrounding area,particularly anteriomedially. The fourth ventricle (4) was displacedanteriorly and to the left. The vermis (V) was also displaced to theopposite side. The posterior fossa cisterns were narrow on the right(unlabelled double arrow). FIG. 4A also shows B and C, which shows apost gadolinium MR section in exaggerated inferior orbitomeatal planetaken on Jun. 16, 1994. Because of considerable difference in sectionalangles between A and B & C, two consecutive MR sections (B &C) wereselected in order to amply cover the original site of the cerebellarmetastasis. The enhancing tumor attenuated and was no longer detectablein post gadolinium axial MR T1 weighted image. Unlabelled single arrowin two consecutive MR images indicates the presumptive location of theoriginal cerebellar metastatic lesion seen in A. The fourth ventricle(4) and the vermis (V) have returned to their normal positions. Theedema surrounding the tumor seen before the treatment was not observed.FIG. 4B is a radiograph of the left scapula of patient #8. In A, theradiograph of left scapula shows a mixed osteoblastic and osteolyticlesion (white arrowhead) in the inferior aspect of the glenoid process,due to metastatic cancer. In B, the radiograph of left scapula 12.5months later shows that the lesion in the glenoid process has muchimproved. Patient #11 had radiotherapy to her left cerebellar brainlesions; her primary tumor and the multiple lesions in her brainstemremained stable without radiotherapy for 10 months.

[0051] Toxicity and Performance Status: Patients ingested SV daily for 5to 46 months. Except for patient #14 whose condition was adverselyaffected by concurrent chemotherapy, all patients in the cohort (#3-#13)showed no toxic complications attributable to SV. Their blood chemistryvalues revealed no compromise in hematological, renal, hepatic, andmetabolic function; they all showed a significant improvement in KPSafter ingesting SV for 5 months (Table 2). Average KPS values increasedfrom 54+12 to 90+15 (p<0.01).

Discussion

[0052] The benefits of fruits and vegetables for cancer prevention hasbeen emphasized by both NCI and ACS and are considered as major causesof the reduction in cancer rates and related deaths. The present reportdescribes the pursuit of an unexpected clinical observation and providesdata suggesting that a mixture of edible plants, specifically selectedfor their antitumor attributes, may benefit patients who already haveadvanced cancer.

[0053] The MST of stage IIIB and IV NSCLC has changed little in recentdecades. Therefore, historical controls can be used to providecomparison for the results observed here. In a review of 8 randomizedtrials of chemotherapy versus supportive care for stage III-IV NSCLC,the MST's of the untreated groups were 5 months or less and 7-8.5 monthsin the chemotherapy groups (4). The MST for the 12 patients who used SVfor 2 months or longer was 33.5 months. The sample size was small, butall patients had advanced disease; moreover, objective responses to SVwere observed in this cohort and in one tumor-bearing mouse.

[0054] NSCLC patients with CNS metastases have a MST of 3-4 months withradiotherapy and complete regression is rare (39-41). Of 4 patients withCNS involvement, three (#2, 9, 10) had complete remission of CNS lesionsand-the fourth (#11) had a partial response after radiotherapy and SVuse. The complete regression of metastatic tumors in both brain and bone(#9), and multiple brain lesions (#2) is unexpected. Recurrence ofmalignant pleural effusions and new metastases are common in stageIII-IV patients. Ten of 14 patients developed no new metastatic sitesduring this study period.

[0055] Patient selection bias is a major consideration in clinicaltrials. It is essential to consider sources of potential bias especiallywhen clinical outcomes differ markedly from historical controls. Fourpatients (#15-18) who ingested SV for less than 2 months were notincluded in the survival analysis; their survival times (3 to 10 months)were within the expected historical range. Other possible bias couldresult from the self selection of highly motivated patients who soughtnew treatments after failing to benefit from conventional therapies andwho were willing to make a commitment to daily SV use often with strongfamily support. Cassileth et al. report a weak link between cancersurvival and social ties, but these authors concluded that clinicalfactors (e.g. stage, KPS, weight loss) but not psychosocial factors(e.g. “hopefilness”) could be considered predictors of survival. Whereaspatient compliance may be related to treatment efficacy, psychologicalfactors alone are unlikely to result in a three-fold increase in MST orin complete tumor regression in patients using SV alone. The inhibitionof tumor growth observed in the mouse model further supports thehypothesis that some combination of ingredients in SV may have directanti-tumor or immune modulating activity.

[0056] Large clinical trials of advanced NSCLC often have a small numberof patients with long survivals. It could be argued that by chance orselection bias such patients were overrepresented in this cohort. Thisis unlikely because of many instances of poor prognostic status andfailed treatment history in this cohort when SV was initiated.Furthermore, complete and partial tumor regression and improved KPSafter using SV alone are objective responses to SV, which are notaffected by patient differences or by patient number. The potentialcauses of long survivorship have received relatively little attention,although such studies may yield clues for improved cancer treatment.Among these possible causes, diet supplements are widely accessible andsome are supported by scientific studies. This approach, which uses aspecific dietary supplement, was ingested by the 2 lead patients andthis cohort.

[0057] Well-known prognostic indicators for survival in NSCLC includestage of disease, KPS, and weight loss. KPS values increased from anaverage of 54+12 to 90+15 in 5 months after SV use. In a prior study,the KPS of stage III and IV NSCLC patients of SV-treated group alsoimproved but it decreased in the untreated control group; similarly theMST also improved significantly (3-fold). Body weight data were notavailable in the current study. In the prior study, however, the weightchange was −2.1+2.3% in the SV-treated group but −11.6+4.9% in thecontrol group. In both studies, blood chemistries revealed no evidenceof toxicity in patients ingesting SV daily for 5-46 months.Significantly improved performance status and absence of toxicity areconcordant with the prolonged survival times of these patients.

[0058] The immunostimulatory and antitumor effects of purifiedbeta-glucans (e.g., lentinan, pachymaran, zymosan, schizophyllan) andpeptidomannan (KS-2) have been well documented. Other studies haveidentified antitumor actions of soybean components, e.g., proteaseinhibitors, inositol hexaphosphate, coumestrol, daidzein, biochanin A,and genistein. Four of these components were quantitatively identifiedin SV; others are reported to be present in the vegetables contained inSV. The most abundant ingredients in SV are soybean, mushroom, and mungbean. Because Purina Lab Chow already contains soy protein, theantitumor activities of mushroom and mung bean were tested individuallyand in combination. Combining mushroom and mung bean extracts producedthe greatest inhibition of tumor growth (82%) in the mouse modelindicating that certain combinations of foods may be more carcinostatic.

[0059] The multiple instances of tumor response coupled with increasedsurvival times, attenuation of disease progression, and consistentimprovement in performance status indicate that SV therapy, as anadjuvant to surgery and radiotherapy, should be evaluated further in alarge, randomized study of NSCLC patients.

Example 2 Phase I/II Study of Stage III and IV Non-Small Cell LungPatients Selection of Patients

[0060] All the stage III and IV NSCLC patients who first attended theLung Clinic, School of Medicine, University of Palacky, from June 1992to January 1994 and met the entry criteria were invited to participatein the study. All eligible patients who did not use SV served as thecontrol group (13 patients). Cooperative group phase II-IIIinvestigation entry criteria were used: ≦70 years of age; ability to eatwell; adequate gastrointestinal, hepatic, renal, and cardiopulmonaryfunction; and Kamofsky performance status (KPS) ≧70. Exclusion criteriawere white blood cells <4,000/ml, platelets <100,000/ml, bilirubin >1.5mg/dl, creatinine >1.5 mg/dl, blood sugar >200 mg/dl, prior malignancy,myocardial infarction, uncontrolled hypertension, arrhythmia, seriousinfection, psychiatric disorder, unreliability, or noncompliance. Inaddition to the entry criteria, prognostic factors for survival,including stage, previous and concurrent therapies, tumor cell type,age, gender, KPS, and body mass index (BMI) were compared in all groups.Twelve patients agreed to ingest SV: six at stage III and IV in thetreatment group (SVG) and five at stage I in the toxicity study group(TG); one patient was first diagnosed as having adenocarcinoma but wasexcluded from analyses because she was later identified as having coloncancer in 1983. All patients were offered standard conventionaltherapies. Patients were followed until death or 24 months from the dateof entry.

[0061] Study Period, Survival Time, and Statistical Analyses

[0062] This study was designed to test the null hypothesis (Ho) that themedian survival times of the stage III and IV patients in the treatmentgroup (SVG) and the control group (CG) will be the same. The alternativehypothesis (H₂) is that the MSTs of the two groups will be different.The study period was 24 months from the starting date (date ofdiagnosis). The survival time was calculated from the starting date tothe date of death or to the cutoff date. The Kaplan-Meier method andlog-rank test were used to calculate and compare the median survivaltimes and the 95% confidence intervals. The Wilcoxon Rank and Sum testwas used to compare mean survival times between the two groups.Student's t-test was used to compare the ages, KPS, and percent weightchange.

[0063] Diagnosis, Staging, and Survival Time

[0064] NSCLC was diagnosed by histopathology and/or cytopathology andreviewed by a pathologist. The size and location of the tumors weredetermined with radiographs by the treating physicians and a radiologistand reviewed by another radiologist. Staging was assigned according tothe American Joint Committee on Lung Cancer. The most conservativeinterpretation was used for analyses.

[0065] Treatment

[0066] All patients were treated with conventional therapies, includingradiation, surgery, and/or chemotherapy, selected by the treatingphysicians independently of the use of SV. SV was included as part ofthe daily diet of the treatment group patients. The intended duration oftreatment with SV was 24 months; participating patients were encouragedto ingest SV soon after diagnosis and for the entire period. Alleligible patients who ingested SV were included in the analyses.

[0067] Thirty grams of freeze-dried SV powder were mixed with one cup ofwater or other soup and ingested daily. Patients were interviewed andexamined monthly, except for four patients (Patients 5, 6, 8, and 10,Table 2) who were unable to keep their clinical appointments. Computedtomography scans were performed at entry and then every six months orsooner if clinically needed. Body weight and KPS were evaluated at thestudy entry and one to three months thereafter. Chest films, spirometry,and blood chemistry were performed at entry and when clinically requiredor bimonthly.

[0068] Nutrition Data

[0069] Random samples of SV were sent to Northeast Laboratories fordetermination of their nutritional values. A 30-g daily serving of SV(dry weight), on the basis of percentage of daily requirement of a2,000-calorie diet contains 11 g (4%) carbohydrate, 11 g (24%) protein,<1 mg fate, 8 g (36%) dietary fiber, <1 mg cholesterol, 102-110 calgross food energy, and 84-90 cal physiological fuel value.

[0070] Toxicity

[0071] The toxicity of SV was evaluated in all patients who had ingestedSV. To minimize the confounding effects of advanced disease andtherapeutic toxicity, five stage I NSCLC patients were invited to takeSV daily. Their clinical status was evaluated monthly, and bloodchemistries were obtained at entry and bimonthly. Clinical signs oftoxicity, such as fatigue, vomiting, nausea, diarrhea, fever, headache,hair loss, mucosal ulcerations, and skin rashes were evaluated. Inaddition, changes in appetite and in the frequency of sexual activityand exercise before and after diagnosis of NSCLC and before and aftertaking SV daily were also evaluated.

[0072] Informed Consent

[0073] The protocol for the clinical trial was approved by the Committeefor Research on Human Subjects at the University of Palacky School ofMedicine. Informed consent was obtained from all patients in thetreatment and toxicity study groups.

Results

[0074] Patient Characteristics

[0075] The control group (CG) consisted of five patients at stage 1V,four at stage IIIB, and four at stage IIIA (Table 2). The age of thepatients was 40-65 years, with an average of 54.3±8.8 years. At entrytheir body mass index (BMI) was 19.5-36.5, with an average of 26.7±5.5,and their KPS was 70-90, with an average of 78.5+8.0. Thirty-one percentof the CG patients were women. In the treatment group (SVG), 6 patientswere eligible: one was a woman, two were at stage IV, three at stageIIIB, and one at stage IIIA (Table 3). Their age was 41-55 years, withan average of 49.2±4.7 years. At entry their BMI was 24-30.7, with anaverage of 27.0±2.7, and their Karnofsky performance scale (KPS) was70-90, with an average of 75+8.4. Student's t-test was used to comparethe age, BMI, and KPS at entry of the SVG patients with those of the CGpatients, yielding p>0.1.

[0076] Five stage I patients were in the toxicity study group (TG)(Table 3). At study entry their age was 41-68 years, with an average of54.6±11.3 years; their BMI was 22.2-27.7, with an average of 24.7±2.4;and their KPS was 70-90, with an average of 78.0±8.4. One other patient(Patient 12, Table 3) had colon cancer in 1983 and was presumed to havemetastatic colon cancer on review and was excluded from the analyses.

[0077] Weight Change

[0078] At entry BMIs of CG, SVG, and TG patients were 26.7±5.5, 27±2.7,and 24.7+2.4, respectively (p>0.1, Student's t-test). None of thepatients was severely underweight, and all BMIs were >19. The bodyweight of nine CG patients was measured one to three months after entry(except for Patient 1), and all lost 3.7-17.7% of their body weight,with an average loss of 11.6±4.9% (Table 2). In the SVG patients who didnot receive chemotherapy, there was no change or a loss of <1.5% of bodyweight within the first seven month after entry; those with concurrentchemotherapy lost 4.8% and 5.2% (Table 3). Similarly, the two CGpatients (Patients 9 and 13, Table 2) who were treated with chemotherapylost more weight (17.7% and 16.4%, respectively) than the average(11.6%). The average weight loss of the SVG patients within the firstseven months after entry was 2.1±2.3%. When the change in body weightbetween the two groups was compared, p<0.01.

[0079] The TG patients either gained or had no change in their weightmeasured 4-12 months after entry, with an average weight gain of4.0±3.9%. The ineligible patient (Patient 12) showed no weight change.

[0080] Performance Status

[0081] Within the first three months after entry, virtually all the CGpatients deteriorated rapidly, i.e., their KPS declined to 40-70, withan average of 55.4±11.3 (p<0.01 compared with KPS at entry) (Table 3).Except for one patient, the condition of the SVG patients improved orremained the same, and their KPS was 60-90, with an average of 80±13(p>0.1 compared with KPS at entry) (Table 4). The KPS of the CG and SVGpatients after entry were compared, yielding p<0.01.

[0082] Four of the SVG patients not receiving chemotherapy showed anincrease or no change in KPS. In the two patients treated withconcurrent chemotherapy, there was a decrease or no change in KPS (Table4). The condition of all TG patients and of the ineligible patient(Patient 12) improved, and all were alive and well at 24 months, withKPS of 90. TABLE 3 Patient Characteristics: No Ingestion of SVs^(a) KPSPatient Stage: After BMI, % Wt Survival, No. Age Gender Cell TypeTNM^(b) Treatments At entry treatment^(c) kg/m² Change^(d) mo 1 65 MSquamous IV: 3.2.1. R 80 60 (3) 27.1 −11.0 (7) 8 2 59 F AdenocarcinomaIV: 2.2.1. R 80 50 (3) 36.5  −3.7 (1) 5 3 54 M Squamous IV: 2.2.1. R 7050 (3) 23.6 −13.2 (2) 4 4 45 M Squamous IV: 1.3.1. R 80 50 (2) 28.0 −7.7 (3) 3 5 46 F Undifferentiated IV: 2.1.1. S, R 70 40 (2) 22.9 NA 36 50 M Squamous IIIB: 4.2.0. R 80 70 (1) 26.1 NA 1 7 56 F Poorlydifferentiated IIIB: 3.3.0. R 80 60 (3) 31.2  −8.2 (1) 7 8 60 M SquamousIIIB: 4.0.0. S, R 70 40 (3) 26.6 NA 4 9 62 M Adenocarcinoma IIIB: 4.1.0.C 90 40 (2) 27.5 −17.4 (2) 4 10 65 M Squamous IIIA: 3.2.0. SC 90 70 (1)21.1 NA 2 11 62 M Squamous IIIA: 3.1.0. R 70 60 (3) 22.5 −17.7 (3) 12 1242 M Adenocarcinoma IIIA: 2.2.0. R 70 60 (1) 36.3  −8.7 (1) 2 13 40 FAdenocarcinoma IIIA: 3.2.0. C, R 90 70 (3) 19.5 −16.4 (3) 7 Mean ± SE54.3 ± 8.8 78.5 ± 8.0 55.4 ± 11.3 26.7 ± 5.5 −11.6 ± 4.9 4.8 ± 3.0

[0083] TABLE 4 Characteristics of Patient Who Ingested SV^(a) KPSPatient Stage and After BMI, % Wt Survival, No. Age Gender Cell TypeTNM^(b) Treatments At entry treatment^(c) kg/m² Change^(d) SV mo moStage III and IV patients for survival study  1 50 M Squamous IV: 2.2.1.R 70 90 (3) 26.1 −1.2 (5) 7 >24  2 49 M Squamous IV: 2.1.1. R 70 80 (3)30.7  0.0 (5) 5 16  3 52 M Adeno- IIIB: 4.1.0. R 80 90 (3) 24.0 −1.3 (7)17 18 carcinoma  4 55 M Squamous IIIB: 2.3.0. C 70 70 (3) 29.4 −4.8 (6)4 9  5 41 F Adeno- IIIB: 4.3.0. C 70 60 (3) 27.3 −5.2 (3) 7 8 carcinoma 6 48 M Adeno- IIIA: 2.2.0. R 90 90 (3) 24.1  0.0 (3) 4 15 carcinomaMean ± SE 49.2 ± 4.7    75 ± 8.4 80 ± 13 27.0 ± 2.7 −2.1 ± 2.3 Stage Ipatients for toxicity study  7^(e) 41 M Squamous I: 2.0.0. S, R 70 90(3) 24.1  2.3 (12) >24 >24  8 63 M Squamous I: 1.0.0. R 90 90 (3) 26.7 0.0 (5) >17 >24  9 68 M Squamous I: 1.0.0. R 80 90 (3) 27.7  7.5(7) >20 >24 10 55 M Large cell I: 2.0.0. R 70 90 (3) 22.2  8.8(8) >20 >24 11 46 M Squamous I: 2.0.0. R 80 90 (3) 22.6  1.3 (4) >18 >24Mean ± SE 54.6 ± 11.3 78.0 ± 8.4 90.0 ± 0.0  24.7 ± 2.4  4.0 ± 3.9Ineligible patient 12^(f) 47 F Adeno- N/A S, C, R 70 90 (3) 31.21.2 >24 >24 carcinoma

[0084] Survival Time

[0085] The CG patients survived from 1 to 12 months, and all died beforethe cutoff date, with an average survival time of 4.8±3.0 months (Table3). The median survival time was four months with a 95% confidenceinterval of three to seven months (FIG. 5). If the worst survivors(Patients 6, 10 and 12) were excluded from evaluation, the remainingpatients survived for 3-12 months, and the median survival time was 4.5months, with a 95% confidence interval of 4-7 months, with an averagesurvival time of 5.7±3.0 months.

[0086] SVG patients ingested SV for 4-17 months. The median survivaltime was 15.5 months, and its 95% confidence interval was 9-18 months,and one patient survived >24 months (FIG. 5). In the log-rank testcomparing median survival times of the SVG and the CG patients(including or excluding Patients 6, 10, and 12), p<0.01, indicating thatthe median survival time of the SVG patients is significantly longerthan that of the CG patients. Thus, H₀ is not supported by the data andrejected, but H₂ is supported and accepted. If Patient 1 was assumed tobe dead at the cutoff date (Table 4), the Wilcoxon Rank and Sum test forsmall sample size could then be used. The mean survival times of the SVGand the CG patients were 15 and 4.8 months (or 5.7 months excluding theworst survivors), respectively, with p<0.01, indicating that the formeris significantly longer than the latter. Again H₀ is not supported bythe data and rejected, but H₂ is supported and accepted.

[0087] One patient (Patient 7) had lesions in both adrenal glands, butno biopsy was performed. He was classified as stage I for toxicityevaluation only. One other patient (Patient 12) had colon cancer in 1983and was presumed to have metastatic colon cancer on review. Bothpatients were alive and well beyond 24 months, but they were notincluded in survival analyses.

[0088] Toxicity Evaluation of Patients Ingesting SV

[0089] All patients who ingested SV (Table 4) showed no clinical signsof toxicity (see Materials and Methods). Their blood chemistry dataindicated that no significant hematologic, renal, or hepatic toxicityoccurred. The five stage I TG patients ingested SV daily for 17-24months (Table 4). All were treated with radiotherapy and/or surgery, butnot chemotherapy. None of these patients showed weight loss, bleeding,or any clinical symptoms of gastrointestinal, neurological, orcardiopulmonary dysfunction. All were alive and well with stable tumorat the end of the 24-month study period. They maintained or resumedroutine activities, including daily full-time work, exercise, and sexualactivity. No clinical signs of toxicity were detected. None developednew tumor during the study period. Some patients reported a feeling offullness or bloatedness after ingesting SV, and some found it difficultto consume 30 g of SV daily. A few patients did not care for the tasteof SV. Nonetheless, all five TG patients ingested SV daily for >17months.

Discussion

[0090] This exploratory study tests the hypothesis that the dailyingestion of a combination of foods that contain different types ofantitumor components may benefit patients with stage III and IV NSCLC.Several vegetables contain molecularly defined components withimmune-modulating or antitumor activity (Table 1). The individualactivities, although well documented, have only modest potencies. Theyhave different mechanisms of action and thus may act synergisticallywhen used together. These antitumor entities are not toxic. Adding themto the daily diet of cancer patients incurs minimal risk.

[0091] Conventional therapies afford only marginal benefit for stage IIBand IV NSCLC. The median survival time of stage IIIB and IV NSCLCpatients is usually short. Thus, adding a dietary supplement to thepatients' daily diet without interfering with conventional therapies wasdeemed ethical, and adequate clinical observation could be achievedwithin 24 months.

[0092] The number of SVG patients is small, but the difference in mediansurvival times between CG and SVG patients is large (3-fold). TheLog-Rank test showed p<0.01 and no overlap in the 95% confidenceintervals of the two median survival times, indicating a significantdifference between SVG and CG patients. Furthermore, using the WilcoxonRank Sum test, which could also be used to compare groups of smallsample size, the mean survival time of the SVG patients is alsostatistically longer than that of CG patients (p<0.01). Moreover, usingeven more conservative evaluation, if the CG patients with the shortestsurvivals (Patients 6, 10, and 12) are excluded from analysis, themedian survival time and mean survival time of CG patients are stillsignificantly different from those of SVG patients (p<0.01).

[0093] Most patients failed to ingest SV for the intended period, butall patients ingested SV for four months or longer. Failure to ingest SVfor the intended period was mostly due to lack of motivation andconfidence in the efficacy of SV, boredom and monotony in taking 30 g ofSV powder daily, taste preference of patients unaccustomed tohigh-fiber, low-fat foods, bloating, fullness, and cancer-relatedanorexia. SVG patients (stage III-IV), except for Patient 3,discontinued SV ingestion earlier than TG patients (stage I), possiblybecause they were weaker or more sensitive to one or more of the factorslisted above. No attempt was made to rank the importance of thesefactors. Because no prior. data on efficacy of using SV for differentdurations were available, all these patients were included in analysesto avoid bias.

[0094] Nonrandomized trials are recommended for phase II studies. Biasin patient selection can be minimized by using the same clinical entrycriteria and prognostic factors for patients in the control and thestudy group. In the present study, tumor cell type, stage, previous andconcurrent therapies, age, gender, KPS, and BMI in the CG and the SVGpatients were comparable. KPS, change in body weight, and extent ofdisease are considered strong survival predictors for cancer patients.These prognostic indicators were comparable in the CG and the SVGpatients at entry but rapidly declined in the CG patients. In contrast,they often improved or remained unchanged, or their decline was lessrapid and severe in SVG patients. All these indicators improved or wereunchanged in all TG patients. Thus, ingestion of SV was associated withmaintenance or improvement in KPS and body weight.

[0095] The median survival time of the CG patients was within theexpected historical range. Median survival time and mean survival weresignificantly longer for SVG than for CG patients. For changes of KPSand body weight, each patient was used as his own control. These changeswithin individual patients are not affected by the difference betweenpatients or by the number of patients and are strong prognosticindicators for patient survival. The rapid declines in these survivalindicators in the CG but not in the SVG patients are consistent with theobserved survival difference between the two groups.

[0096] Quality of life and avoidance of drug toxicity, cachexia, andrecurrent and metastatic disease remain important challenges in thetreatment of NSCLC. In the 24-month study period, no detectable toxicityoccurred in the five stage I patients, and their KPS and body weightalso improved. Moreover, although recurrent tumor in stage I NSCLCpatients is a major cause of death, all five stage I TG patients had norecurrent tumor in the two-year study period. They all had primaryradiotherapy, and one also had surgery. Their 24-month recurrence-freesurvival compares favorably with the 59% recurrence rate and the 52%death rate due to lung cancer in two years for stage I NSCLC patientstreated with radiotherapy alone or the 35% two-year recurrence rate ofstage I NSCLC patients treated with surgery alone.

[0097] The apparent benefit of SV may be due to one or more antitumorcomponents in SV (Table 1). The antitumor effect of purified forms ofβ-glucans has been well documented and is host mediated. Soybeanscontain various antitumor components, e.g., protease inhibitors,saponins, inositol hexaphosphate, genistein, and other phytoestrogens.Moreover, NSCLC is an immune-suppressing disease, and immunosuppressivefactors produced by NSCLC cells have been well characterized. Furtherinvestigations of the benefits of SV and other combinations of foodscontaining different antitumor and immune-enhancing entities for cancerpatients are warranted. This approach is attractive because of itssimplicity, low cost, and lack of toxicity.

[0098] This study indicates that the daily ingestion of SV is safe,nontoxic, and well tolerated by NSCLC patient at all stages. Theprolonged survival of stage III/IV NSCLC patients observed in this phaseI/II study is noteworthy and warrants further large, placebo-controlled,randomized studies. Given the high incidence and poor prognosis of lungcancer, modest improvements in survival can be translated into manythousands of useful added years for NSCLC patients.

[0099] Furthermore, strong Antitumor efficacy of SV was also observed in2 tumor models in mice, namely, lung carcinoma Line I in BALB/c mice andSarcoma S1509a in A/J(H-2a) mice (2,3).

Example 2 Colon Cancer

[0100] Subsequently, a Czech patient with stage 1V colon cancer had anunusually prolonged survival (>4 years) with conventional therapies anddaily ingestion of SV.

[0101] Four sets of CT scans were taken on the following dates: Oct. 13,1986, Nov. 11, 1986, Nov. 26, 1986, and Feb. 10, 1987. Threeintracranial enhancing lesions (FIGS. 2A, 2B, and 2C) were identified;their transverse directions were measured (in mm).

[0102] In FIG. 2A, the lesion, located in the left posterio-lateralmiddle frontal gyrus, was surrounded by edema anteriorly and medially.No obvious focal indentation, deformity, or shift of the ventricularsystem was observed. The lesion measured 13 mm (A1); 13.6 mm (A2); 8.6mm (A3); no clear cut enhancing mass lesion was identified in the area(A4). The edema represented by areas of low density shows markeddiminution. Unlike the first, second, and third CT scans, where thelesions are still clearly seen, the last CT scan was taken with a higherquality scanner (GE 9800 scanner). Therefore, even a small enhancinglesion, if still present, should be better demonstrated.

[0103] In FIG. 2B, the lesion, located in the medial aspect of theprecentral gyrus on the left side immediately adjacent to the falx,measured 7.7 mm (B1); 11.5 mm (B2); 6.7 mm (B3); no clear cut enhancingmass lesion was identified in the area (B4).

[0104] In FIG. 2C, the lesion was located in the precentral gyrus (upperconvexity side). A moderate degree of edema was seen involving the whitematter of the high frontoparietal lobe. The pre- and postcentral gyriwere widened and the adjacent sulci were narrowed. This lesion measured11.5 mm (C1); 12.1 mm (C2); 6.0 mm (C3); no clear cut enhancing masslesion was identified in the area (C4).

Example 3 AIDS Treatment

[0105] An AIDS patient was administered SV when his CD4⁺ decreased to adangerously low level (95 cells/mm³) condition deteriorated, and hadmultiple infections. After ingesting SV for 2 months, not only hishealth improved to normal level, his CD4⁺ cell count also increased tonormal level, namely, to 475 in 2 months and over 600 in 6 months. Thispatient did not receive any other treatment except for daily ingestionof SV during this time. Thus, the change in his CD4⁺ cell count andhealth condition after ingesting SV alone is considered an objectiveresponse to SV.

1. A method to ameliorate at least one effect of malignancy or viralinfection which method comprises: administering to a subject in need ofsuch treatment an effective amount of a dietary supplement comprisingsoybean, mushroom and mung bean or extracts thereof, wherein saidcomposition is administered periodically for a time sufficient toachieve amelioration.
 2. The method defined in claim 1 wherein saiddietary supplement further comprises at least one member of the groupconsisting of red date, scallion, garlic, lentil bean, leek, hawthornfruit, onion, ginseng, angelica root, licorice, dandelion root, senegalroot, ginger, olive, sesame seed, and parsley, or extracts thereof.
 3. Amethod of enhancing immune functions comprising administering to asubject an effective amount of a dietary supplement comprising soybean,mushroom and mung bean or extracts thereof administered periodically fora time to achieve enhanced immune functions.
 4. The method defined inclaim 3 wherein said dietary supplement further comprises at least onemember of the group consisting of red date, scallion, garlic, lentilbean, leek, hawthorn fruit, onion, ginseng, angelica root, licorice,dandelion root, senegal root, ginger, olive, sesame seed, and parsley,or extracts thereof.
 5. The method defined in claim 1 wherein saidmethod increases CD4⁺ cell levels in said subject.
 6. The method definedin claim 5 wherein said dietary supplement further comprises at leastone member of the group consisting of red date, scallion, garlic, lentilbean, leek, hawthorn fruit, onion, ginseng, angelica root, licorice,dandelion root, senegal root, ginger, olive, sesame seed, and parsley,or extracts thereof.
 7. The method defined in claim 1 wherein saidmalignancy is selected from the group consisting of non-small cell lungcancer, small cell lung cancer, prostate cancer, breast cancer, stomachcancer, small intestine cancer, colonrectal cancer, renal cellcarcinoma, soft tissue sarcoma, ovarian cancer, thyroid cancer,acrcinoid tumors, sarcoma of the bone, melanoma, glioblastoma,oligodendroglioma, astrocytoma, medulloblastoma, acute lymphomas,chronic leukemias, cancer of unknown primary site, Hodgkin's disease,and non-Hodgkin's disease.
 8. The method defined in claim 1 wherein saidviral infection is selected from the group consisting of HIV,Epstein-Barr virus, herpes, and influenza viruses.
 9. A method toameliorate at least one effect of malignancy or viral infectioncomprising administering to a subject in need thereof an effectiveamount of a dietary supplement comprising at least one phytoestrogen, atleast one β-glucan, at least one saponin, inositol hexaphosphate, and atleast one lectin.
 10. The method defined in claim 9 wherein the at leastone phytoestrogen is at least one member selected from the groupconsisting of genistein, coumestrol, daidzein, and biochanin A; whereinsaid at least one β-glucan is at least one member selected from thegroup consisting of lentinan, pachymaran, zymosan, schizophyllan, KS-2,and pachyman; and said at least one lectin is at least one memberselected from the group consisting of phytohemagglutinin andconcanavalin A.
 11. The method defined in claim 10 wherein the dietarysupplement comprises genistein, biochanin A, coumestrol, inositolhexaphosphate, at least one beta-glucan and at least one lectin.